Characterization of Passionfruit severe leaf distortion virus,a novel begomovirus infecting passionfruit in Brazil,reveals a close relationship with tomato‐infecting begomoviruses

Molecular and biological characterization of the begomovirus isolate BR:LNS2:Pas:01, obtained from yellow passionfruit plants in Livramento de Nossa Senhora, Bahia state, Brazil, was carried out. Sequence analysis demonstrated that the BR:LNS2:Pas:01 DNA‐A had highest nucleotide sequence identity with Tomato chlorotic mottle virus (77%) and had five ORFs corresponding to the genes cp, rep, trap, ren and ac4. The DNA‐B had highest nucleotide sequence identity with Tomato yellow spot virus (74%) and two ORFs corresponding to the genes mp and nsp. These identity values indicate that this isolate represents a new begomovirus species, for which the name Passionfruit severe leaf distortion virus (PSLDV), is proposed. Phylogenetic analysis clustered the PSLDV DNA‐A and ‐B in a monophyletic branch with Brazilian tomato‐infecting begomoviruses. The isolate’s host range was restricted to species from the Passifloraceae and Solanaceae. PSLDV‐[BR:LNS2:Pas:01] was capable of forming pseudorecombinants with tomato‐infecting begomoviruses, reinforcing its close relationship with these viruses and suggesting a possible common origin. However, the virus was not capable of infecting tomato.     

Genetic analysis and molecular mapping of quantitative trait loci in common bean against Pythium ultimum

Pythium ultimum is a soil pathogen that can cause seed decay and damage to roots in common bean. In this study, the response of a set of 40 common bean genotypes to P. ultimum and inheritance of the resistance in the 92 F? recombinant inbred lines (RIL) developed from a cross between Xana and Cornell 49242 was investigated by using emergence rate and seedling vigor. Emergence of the 40 genotypes showed a significant association between white seed coat and response to this pathogen. Among these, 11 common bean genotypes, all with colored seeds, exhibited a high percentage of emergence and seedling vigor not significantly different (P > 0.05) to noninoculated plants. Response of the RIL population revealed both qualitative and quantitative modes of inheritance. A major gene (Py-1) controlling the emergence rate was mapped in the region of the gene P, a basic color gene involved in control of seed coat color, located on LG 7. Using the RIL subpopulation with colored seeds, a significant quantitative trait loci (QTL) associated with the emergence rate (ER3(XC)) and another with seedling vigor (SV6(XC)) were identified on the LG 3 and 6, respectively. QTL SV6(XC) was mapped in the region of the gene V, another gene involved the genetic control of color. QTLs associated with seed traits were mapped in the same relative position as regions involved in responses to P. ultimum suggesting the possible implication of avoidance mechanisms in the response to this pathogen.     

Diversity of Campylobacter jejuni and Campylobacter coli Genotypes from Human and Animal Sources from Rio de Janeiro, Brazil

To compare the genotypes of Campylobacter jejuni and Campylobacter coli isolates of human and animal origin collected in Rio de Janeiro City, 30 C. jejuni and 35 C. coli isolates from animal sources (n = 45) and human patients with gastroenteritis (n = 20) were genotyped by PCR-based techniques, namely random amplified polymorphic DNA (RAPD-PCR) and enterobacterial repetitive intergenic consensus sequence (ERIC-PCR). RAPD-PCR identified 50 types and ERIC-PCR identified 22 genotypes, among the 65 Campylobacter isolates. Both PCR methods discriminated the C. jejuni and C. coli groups of isolates. Combining the results of both methods, no single genotype was shared between isolates from human and animal sources. Two groups of two C. coli isolates each with identical genotypes were found among poultry and pig isolates. A high level of genetic diversity observed among the Campylobacter isolates suggests lack of overlap between isolates from different sources.     

Early acute depletion of lymphocytes in calicivirus-infected adult rabbits

Rabbit Haemorrhagic Disease (RHD) is a lethal infection caused by calicivirus that kills 90% of the infected adult rabbits within 3 days. The calicivirus replicates in the liver and causes a fulminant hepatitis. Most studies on the pathology of RHD have been focused on the fulminant liver disease. This may not be the only mechanism in the pathogenesis of RHD: calicivirus infection may also induce leukopenia in the infected adult rabbits. We show now by flow cytometry analysis that the calicivirus induces an early decrease in B and T cells, in both spleen and liver. The depletion of B and T cells was associated with apoptosis labelled by annexin V. These changes occurred in rabbits before they showed enzymatic evidence of liver damage and persisted after liver transaminase values were very high. We conclude that depletion of lymphocytes caused by the calicivirus infection precedes or attends liver damage. The relative contribution of this lymphocyte depletion for the pathogenesis of the fatal calicivirus infection of rabbits remains to be investigated.     

Intraspecific variation in 16S rRNA gene of Mycoplasma synoviae determined by DNA sequencing

Mycoplasma synoviae (MS) is an important avian pathogen may cause both respiratory disease and joint inflammation synovitis in poultry, causing economic losses to the Brazilian poultry industry. The genotypic variation in 16S rRNA gene is unknown. Partial sequences of 16S rRNA gene of 19 strains of M. synoviae were sequenced and analyzed in order to obtain molecular characterization and evaluation of the genetic variability of strains from distinct Brazilian areas of poultry production. Different polymorphic patterns were observed. The number of polymorphic alterations in the studied strains ranged from 0 to 6. The nucleotide variations, including deletion, insertion and substitutions, ranged from 3 to 5. The genotypic diversity observed in this study may be explained by spontaneous mutations that may occur when a lineage remains in the same flock for long periods. The culling and reposition in poultry flocks may be responsible for the entry of new strains in different areas.     

Effects of methadone on the minimum alveolar concentration of isoflurane in dogs

ObjectiveTo investigate the effects of methadone on the minimum alveolar concentration of isoflurane (ISO_MAC) in dogs.Study designProspective, randomized cross-over experimental study.AnimalsSix adult mongrel dogs, four males and two females, weighing 22.8 ± 6.6 kg.MethodsAnimals were anesthetized with isoflurane and mechanically ventilated on three separate days, at least 1 week apart. Core temperature was maintained between 37.5 and 38.5 °C during ISO_MAC determinations. On each study day, ISO_MAC was determined using electrical stimulation of the antebrachium (50 V, 50 Hz, 10 mseconds) at 2.5 and 5 hours after intravenous injection of physiological saline (control) or one of two doses of methadone (0.5 or 1.0 mg kg^?1).ResultsMean (±SD) ISO_MAC in the control treatment was 1.19 ± 0.15% and 1.18 ± 0.15% at 2.5 and 5 hours, respectively. The 1.0 mg kg^?1 dose of methadone reduced ISO_MAC by 48% (2.5 hours) and by 30% (5 hours), whereas the 0.5 mg kg^?1 dose caused smaller reductions in ISO_MAC (35% and 15% reductions at 2.5 and 5 hours, respectively). Both doses of methadone decreased heart rate (HR), but the 1.0 mg kg^?1 dose was associated with greater negative chronotropic actions (HR 37% lower than control) and mild metabolic acidosis at 2.5 hours. Mean arterial pressure increased in the MET1.0 treatment (13% higher than control) at 2.5 hours.Conclusions and clinical relevanceMethadone reduces ISO_MAC in a dose-related fashion and this effect is lessened over time. Although the isoflurane sparing effect of the 0.5 mg kg^?1 dose of methadone was smaller in comparison to the 1.0 mg kg^?1 dose, the lower dose is recommended for clinical use because it results in less evidence of cardiovascular impairment.     

Assessing insecticide and fungicide effects on the culturable soil bacterial community by analyses of variance of their DGGE fingerprinting data

To assess the effects of three insecticides (aldicarb, chlorpyrifos, deltamethrin) and two fungicides (tebuconazole and metalaxyl + mancozeb) on the PCR-DGGE fingerprints of culturable soil bacterial communities (CSBC), a greenhouse experiment was carried out with soil samples from an Integrated System for Agroecological Production (ISAP), a Conventional Potato Production Area (CPPA) and a Secondary Forest Area (SFA) close to the CPPA. Samples were obtained at 15 day intervals starting at 32 until 77 days after sowing (DAS) to perform the PCR-DGGE analysis of the CSBC cultured on media amended with soil suspension. Analysis of variance from PCR-DGGE data indicated significant differences among treatments. Regardless the type of pesticide applied, CSBC was disturbed and similarity values varied from 5% to 90% in comparison to the control. Significant shifts on CSBC were only detected among treatments in the first two harvests, while CSBC tended to be more akin to each other at the last two harvest dates. The most significant responses observed were due to different soil sample origins, where values of 5% of similarity to the control were observed on CPPA soil. The use of analysis of variance on PCR-DGGE data was useful to a better understanding of the changes on CSBC induced by pesticides applications.